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Table of Contents
Chapter 1: Introduction...................................................................................................................4
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B.
C.
D.
User Manual...................................................................................................................................... 4
Purpose of Manual............................................................................................................................ 4
Intended Users.................................................................................................................................. 4
Customer Service and Technical Support ......................................................................................... 4
Chapter 2: Overview of Conjugation ..............................................................................................5
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B.
C.
D.
E.
Product Description .......................................................................................................................... 5
All-in-OneTM Conjugation Technology............................................................................................... 5
All-in-OneTM Conjugation Process Summary..................................................................................... 8
Materials Provided and Storage Conditions ..................................................................................... 9
Additional Materials Required But Not Provided ............................................................................. 9
Chapter 3: All-in-OneTM Conjugation Protocol...............................................................................9
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B.
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D.
E.
F.
G.
H.
I.
IgG Sample Preparation (5 minutes)............................................................................................... 10
Buffer Exchange IgG (3 minutes)..................................................................................................... 10
HyNic Modify IgG (2 h).................................................................................................................... 12
Buffer Exchange IgG (3 minutes)..................................................................................................... 12
Conjugate Formation (2 h).............................................................................................................. 13
Conjugate Purification Stage 1: Affinity Isolation (1 ¾ hours) ........................................................ 13
Buffer Exchange Conjugate (3 minutes) ......................................................................................... 15
Conjugate Purification Stage 2: Q Spin Filter (35 min) ................................................................... 16
Buffer Exchange Conjugate (3 minutes) ......................................................................................... 17
Chapter 4: Appendix......................................................................................................................18
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B.
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D.
E.
F.
G.
H.
Human IgG-R-PE Conjugate (Monoclonal): An example................................................................ 18
Bradford Protein Assay ................................................................................................................... 19
Using a NanoDropTM to Measure Antibody Concentration ............................................................ 21
R-PE Absorption Spectrum (Unmodified R-PE)............................................................................... 24
R-PE Absorption Spectrum (4FB-modified)..................................................................................... 25
Concentration of Dilute Antibody Solutions ................................................................................... 25
Troubleshooting Guide ................................................................................................................... 27
References ...................................................................................................................................... 28
Catalog # A-9001-006
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